Mutational analysis of the IgE-binding epitopes of P34/Gly m Bd 30K

Ricki M. Helm, Gael Cockrell, Cathie Connaughton, C. Michael West, Eliot Herman, Hugh A. Sampson, Gary A. Bannon, A. Wesley Burks

Research output: Contribution to journalArticle

68 Scopus citations

Abstract

Background: Peanuts and soybeans are 2 foods that have been shown to be responsible for many atopic disorders. Because of their nutritional benefit, soybean proteins are now being used increasingly in a number of food products. Previous studies have documented multiple allergens in soybean extracts, including glycinin, β-conglycinin, and the P34/Gly m Bd 30K protein. Objective: Our overall goal was to identify soybean-specific allergens to begin to understand molecular and immunochemical characteristics of legume proteins. The specific aim of the current investigation was to identify the essential amino acid residues necessary for IgE binding in the 5 distinct immunodominant epitopes of P34/Gly m Bd 30K. Methods: Serum IgE from 6 clinically sensitive soybean-allergic individuals was used to identify P34/Gly m Bd 30K in the native and single amino acid substituted peptides with use of the SPOTS peptide synthesis technique to determine critical amino acids required for IgE binding. Results: The intensity of IgE binding and epitope recognition by serum IgE from the individuals varied substantially. With use of serum from 6 clinically soybean-sensitive individuals, 2 of the 5 immunodominant epitopes could be mutagenized to non-IgE binding peptides. Conclusions: Single-site amino acid substitution of the 5 immunodominant epitopes of Gly m Bd 30K with alanine revealed that IgE binding could be reduced or eliminated in eoitopes 6 and 16 in the serum obtained from 6 soybean-sensi-tive patients.

Original languageEnglish (US)
Pages (from-to)378-384
Number of pages7
JournalJournal of Allergy and Clinical Immunology
Volume105
Issue number2 II
DOIs
StatePublished - 2000

Keywords

  • B-cell epitopes
  • P34/Gly m Bd 30K
  • Soybean allergen

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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    Helm, R. M., Cockrell, G., Connaughton, C., West, C. M., Herman, E., Sampson, H. A., Bannon, G. A., & Burks, A. W. (2000). Mutational analysis of the IgE-binding epitopes of P34/Gly m Bd 30K. Journal of Allergy and Clinical Immunology, 105(2 II), 378-384. https://doi.org/10.1016/s0091-6749(00)90091-5